Reproducibility of 10x Genomics single cell RNA sequencing method in the immune cell environment
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
Abstract
10x Genomics is a highly accessible single cell RNA sequencing platform that allows for simultaneous gene expression analysis and identification of receptor chain combinations in cells of the adaptive immune system. Here, we asked whether the gene and receptor expression measurements in peripheral blood mononuclear cells (PBMC) are influenced by technical, cell freezing, FACS-processing, and day to day biological variation. No differentially expressed gene was observed between 1. triplicates aliquots taken from the same vial of frozen PBMC; 2. triplicate vials of frozen PBMC; and 3. triplicate aliquots taken from the same vial of frozen PBMC and processed separately for FACS staining and sorting of different PBMC populations. A small number of differentially expressed genes were observed between PBMC sampled, isolated and frozen from the same donor on different days, and these differences were more pronounced in the memory B cells than other cell populations. T cell receptors were recovered in all replicates when at least 5 cells per clonotype were identified. These findings show high reproducibility of 10x Genomics single cell RNA sequencing data in the immune cell context.
Details
Original language | English |
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Article number | 113227 |
Journal | Journal of immunological methods |
Volume | 2022 |
Issue number | 502 |
Publication status | Published - 1 Mar 2022 |
Peer-reviewed | Yes |
External IDs
PubMed | 35031279 |
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unpaywall | 10.1016/j.jim.2022.113227 |
Mendeley | 7cea6be2-d3df-3c6a-bf8a-4a209a9f931e |
ORCID | /0000-0002-8704-4713/work/141544357 |
ORCID | /0000-0001-9599-8632/work/142241744 |
Keywords
Research priority areas of TU Dresden
DFG Classification of Subject Areas according to Review Boards
Subject groups, research areas, subject areas according to Destatis
Sustainable Development Goals
ASJC Scopus subject areas
Keywords
- 10x genomics, Gene-expression analysis, Peripheral blood mononuclear cells (PBMC), Reproducibility, Single-cell, T cell receptor (TCR), Reproducibility of Results, Genomics, Leukocytes, Mononuclear/metabolism, Sequence Analysis, RNA/methods